Methods and compositions for increasing sex steroids and growth hormones

ABSTRACT

Embodiments of the invention generally relate to methods and supplements for increasing sex steroids and human growth hormone in a human being.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation/divisional of U.S. patent applicationSer. No. 13/623,105, filed Sep. 19, 2012, now U.S. Pat. No. 8,747,922issued on Jun. 10, 2014, the disclosure of which is hereby incorporatedherein in its entirety by this reference.

FIELD OF THE DISCLOSURE

The present disclosure generally relates to methods and supplements forimproving health in a subject. In some embodiments, a nutritionalsupplement may be administered to a subject to increase levels of growthhormone (GH) and sex hormone (e.g., testosterone) in the subject.

BACKGROUND

The primary biological function of human growth hormone (hGH) includesstimulating growth, cell repair and regeneration. Once the primarygrowth period of adolescence concludes, the primary function of hGH inadulthood becomes that of cell regeneration and repair, helpingregenerate skin, bones, heart, lungs, liver and kidneys to theiroptimal, youthful cell levels. As is the case with many of our otherhormones or their pre-cursors, such as testosterone, estrogen,progesterone, DHEA and melatonin, hGH levels decline with age.Therapeutically, many of these hormones can be replaced to offset someof the effects of aging such as menopausal symptoms in women or erectiledysfunction in men.

The human body, like every other living entity, works on daily, orcircadian, as well as monthly and annual rhythms. Daily growth hormonesecretion diminishes with age with roughly half the levels at age fortythat we had when we were twenty, and about one-third of those youthfullevels at age sixty. In some sixty-year olds, the levels are as low as25% of the hGH levels in a twenty-year old. Symptoms of aging includeloss of muscle, increase of fat, decreased physical mobility, decreasedenergy levels and as a result, diminished socialization, diminishedhealing ability and an increased risk of cardiovascular disease anddecreased life expectancy. Low hGH levels are associated with the agingprocess and early onset of disease. For example, Rosen and Bengtssonnoted an increased death rate from cardiovascular disease in hGHdeficient patients. (Rosen, T., Bengtsson, B. A., Lancet 336 (1990):285-2880)

Until recently, hGH was available only in expensive injectable forms,and benefits from the restoration of hGH levels available only to thosewith the ability to pay. Most recently substances that can trigger therelease of human growth hormone from an individual's own anteriorpituitary gland have become available. These are generically referred toas secretagogues. Secretagogues have the ability to restore hGH levels,potentially to the levels found in youth. See, e.g., “Grow Young WithhGH” by Dr. Ronald Klatz, President of the American Academy ofAnti-Aging, published in 1997 by Harper Collins.

HGH-deficient adults have marked reductions in lean body mass, andwithin months of hGH treatment, gains in lean body mass, skin thicknessand muscle mass are observed. (Cuneo R C et al. J Appl Physiol 1991;70:695-700; Cuneo R C et al. J Appl Physiol 1991; 70:688-694; Rudman Det al. N Engl J Med 1969; 280:1434-1438).

It is well-established that intravenous (IV) administration of someamino acids results in significant hGH secretion. Intravenous infusionof 183 mg of arginine/kg body weight in females increased hGHlevels >20-fold and 30 g of arginine elevated serum hGH levels 8.6 foldin males (Merimee T J et al. N Engl J Med 1969; 280:1434-1438; Alba-RothJ et al. J Clin Endocrinol Metab 1988; 67:1186-1189). Other amino acids,such as methionine, phenylalanine, lysine, histidine, and ornithine havealso led to marked increases in hGH (Alba-Roth, Muller, Schopohl, & vonWerder, 1988; Chromiak & Antonio, 2002; Gourmelen, M., M. Donnadieu, etal. (1972) Ann Endocrinol (Paris) 33(5): 526-528).

Given the difficulties in IV administration of amino acids forwidespread use, interest in elucidating the hGH response to oral aminoacid supplements prompted testing of such supplements containing mainlyarginine, lysine and ornithine at varying amounts. Yet the pronouncedvariability in results among these studies make clear the complexitiesinvolved in the design of an effective supplement for supporting hGHlevels in the general public. (Suminski R R et al. Int J Sport Nutr1997; 7:48-60; Lambert M I et al. Int J Sport Nutr 1993; 3:298-305;Corpas E et al. J Gerontol 1993; 48:M128-M133; Isidori A et al. Curr MedRes Opin 1981; 7:475-481; Fogelholm G M et al. Int J Sport Nutr 1993;3:290-297; Chromiak J A, Antonio J. Nutrition 2002 July;18(7-8):657-61).

Testosterone is a steroid hormone synthesized from cholesterol andoccurs in both bound and unbound (free) forms in the body. (Feldman etal., 2002) Though present in small amounts in females, this hormonedetermines sexuality in men and hence is considered the male sexhormone. Low testosterone levels in men can have adverse effects such asincomplete sexual development, reduced libido, decreased muscle mass andstrength, loss of body hair, gynecomastia (the term for abnormal mammarygland development in men relating to breast enlargement), impairedspermatogenesis, and underdeveloped testes. (Salenave, Trabado, Maione,Brailly-Tabard, & Young, 2012) The levels of testosterone in men areshown to decrease with increasing age. Total testosterone levels aresaid to decline at a rate of 1.6% per year, and levels of bioavailabletestosterone are reduced by approximately 2-3% per year according to thelongitudinal results obtained from the Massachusetts Male Aging Study.(Feldman et al., 2002).

Serum hGH levels differ in relation to various factors including age,gender, hormone status, and BMI. (Iranmanesh, Lizarralde, & Veldhuis,1991); (Chowen, Frago, & Argente, 2004) The effect of the sex hormoneson hGH becomes clear in the maturation process most obviously atpuberty, wherein sex hormones have both organizational and activationaleffects related to hGH synthesis. ((Chowen et al., 2004) Testosteronetreatment in testosterone-deficient men has been shown to increase hGHsecretion. (Liu, Merriam, & Sherins, 1987) Furthermore, testosteroneadministration in normal men also increases growth hormone releasinghormone (GHRH) induced hGH secretion, (Devesa et al., 1991; Hobbs,Plymate, Rosen, & Adler, 1993) signifying the association between theendocrine axes of these two hormones.

However, the locus and mechanism by which sex steroids contribute to themodulation of GH secretion in humans is not clearly understood. Devesaet al. (1991) J. Steroid Biochem. Mol. Biol. 40(1-3):165-73. Secretionof GH from the pituitary gland is episodic, and dependent upon, interalia, the episodic release of growth hormone releasing hormone (GHRH)and somatostatin. Id. GH secretion may be determined by a balance ofcontradictory effects of different sex hormones. For example, inhibitoryeffects of 17β-estradiol and stimulatory effects of testosterone withregard to hypothalamic somatostatin release may act in concert tomodulate GH secretion. Id.

BRIEF SUMMARY OF THE DISCLOSURE

Described herein are nutritional supplements and methods for using thesame. In embodiments, the nutritional supplement may be an amino acidsecretagogue composition, which, when administered orally, stimulatesthe pituitary gland of a subject to release growth hormone (e.g., hGH),and results in the release of testosterone from the testes.

Some embodiments include an oral nutritional supplement that comprises,for example and without limitation: L-arginine, Oxo-proline, L-lysine,and cysteine. Particular examples include an oral nutritional supplementthat consists essentially of L-arginine hydrochloride, Oxo-proline,L-lysine hydrochloride, N-acetyl-L-cysteine, L-glutamine, andschizonepeta powder.

Certain embodiments herein include an oral nutritional supplement dosageform that consists of 0.86 mmol L-arginine; 1.32 mmol Oxo-proline; 2.05mmol L-lysine; 1.53 μmol N-acetyl L-cysteine; 1.71 μmol L-glutamine; and125 μg Schizonepta (aerial parts) powder. This oral nutritionalsupplement is referred to herein as “SeroVital™.” SeroVital™ may beorally administered in an amount of, for example, 2.9 grams (i.e., 4unit dosage forms) to a human being, so as to stimulate the release ofhGH and/or increase testosterone levels in the human being. Thenutritional supplement may be administered on a regular basis, such as aweekly or monthly intake at a dosage tailored to the subject's needs;i.e., the nutritional supplement may be administered regularly asmultiples (1×, 2×, etc.) of the structural units (pills, tablets,capsules, etc.) in accordance with the needs of the subject.

Some embodiments include a method for jointly increasing growth hormone(e.g., hGH) and androgen (e.g., testosterone) levels in a subject (e.g.,a human subject) that comprises administering (e.g., orally) anutritional supplement to a subject to improve one or more objectivehealth metrics, including for example and without limitation: increasinglean body mass; reducing obesity, adipose tissue mass, and anxiety;supporting weight loss; decreasing appetite and atrophic processes inskeletal muscle, liver, kidney, spleen, skin, and bone; and improving atleast one of energy, endurance, sleep, metabolism, heart rate, bloodpressure, cardiovascular health, sympathetic nervous activity, thyroidresponse, glucose utilization, mental/cognitive function, reaction time,bone density, hair health and appearance, nail health and appearance,skin health and appearance, libido, and at least one androgen-mediatedcondition(s).

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 includes a plot of growth hormone levels measured in subjectsafter administration of an exemplary supplement compared to a placebo;

FIG. 2 includes a scatter plot and linear regression analysis of thetime to fall asleep in subjects with continued use of an exemplarysupplement over time; and

FIG. 3 includes a scatter plot and linear regression analysis of thetime awake during sleep over time in subjects with continued use of anexemplary supplement.

DETAILED DESCRIPTION

The determination of an effective and safe oral functional blend thatstimulates hGH secretion in the general population is important, sinceathletes, entertainers, and now the general public, seek effective hGHsupport supplements and understand hGH to have rejuvenating properties.Indeed, once partial to athletes and entertainers, the desire foreffective supplements to provoke hGH increases now extends to thegeneral public. Not only do they have a goal of building lean tissue andreducing fat, but also of improving skin quality and providing otherrejuvenating qualities that hGH is understood to provide. However, theliterature on oral amino acids for use in stimulating hGH does notcontain clear evidence for an optimized oral amino acid-containing blendable to stimulate hGH in the general public, including both men andwomen of a wide age range.

The original and primary therapeutic use of testosterone was for thetreatment of men with low levels endogenous testosterone production.According to this use, serum testosterone levels are brought to andmaintained in a normal range. However, testosterone may also bedesirably increased in a subject for one of many other conditions andpurposes besides replacement. For example and without limitation,testosterone therapy may be desired to reduce infertility; increaselibido and/or correct erectile dysfunction; treat osteoporosis;encourage penile enlargement; encourage height growth; encourage bonemarrow stimulation; reversing the effects of anemia; and increaseappetite. Testosterone therapy may be particularly desirable in agingindividuals exhibiting decreased androgen production.

Embodiments herein provide a nutritional supplement for jointlyelevating growth hormone (e.g., hGH) and testosterone release.Particular embodiments provide an amino acid-containing composition thatis well tolerated, and may have the result of increasing or elevatinghGH release in those individuals whose hGH release rates have slowed asa function of increasing age, or that have normal hGH levels but desirehigher hGH levels. Compositions may also have the result of increasingor elevating testosterone levels. There is a need for a nutritionalsupplement that efficiently and demonstrably enhances the production andeffects of natural human growth hormone and testosterone individuals ofthe general population.

Some embodiments herein provide a nutritional supplement for use by ahuman being. In particular embodiments, the nutritional supplement is anamino acid secretagogue composition, which, when administered orally,may stimulate the pituitary gland to produce hGH, and may jointlystimulate the testes to produce testosterone.

Increased production of hGH may result in inhibition of insulindepression; inhibition of hyperglycemia and increase in insulineffectiveness; enhancement of fat conversion; lowering of cholesterol;and/or normalization of lipid balance. Increased production oftestosterone may result in modulation of physiological or pathologicalconditions mediated by androgens and the interaction of androgens withtheir receptors. In examples, a supplement herein may function as adietary supplement by assisting the body's own ability to secrete hGHand testosterone naturally, and in a manner which is safe and effective.Such a supplement may provide growth hormone therapy and testosteronetherapy in a more affordable manner than existing compositions andmethods, for example, injectable steroid compositions.

Particular embodiments herein include an oral nutritional supplementthat comprises L-lysine, L-arginine, oxo-proline, and one of cysteineand glutamine. In some examples, a supplement herein may comprise bothcysteine and glutamine and/or schizonepeta powder. In particularexamples, a functional dosage includes the L-arginine at a level betweenabout 0.1-6 mmol and the oxo-proline between about 0.1-8 mmol, and/orthe L-lysine in an amount between about 0.1-12 mmol. In particularexamples, the cysteine and/or glutamine may be contained at a levelbetween about 0.001-6 mmol.

Cysteine may be present in a supplement according to particularembodiments as n-acetyl L-cysteine, and the glutamine may beL-glutamine. Amino acids in a nutritional supplement herein may bedelivered as non-toxic salts thereof, effective complexes thereof,stable chelates thereof, active esters thereof, functional derivativesthereof, and mixtures thereof which are effective to increase hGH levelsin a subject from the general population.

Particular embodiments herein include an oral nutritional supplementthat consists essentially of L-lysine (e.g., L-lysine HCl), L-arginine(e.g., L-arginine HCl), oxo-proline, N-acetyl-L-cysteine, L-glutamine,and schizonepeta (aerial parts) powder. In particular examples, afunctional dosage includes L-arginine at a level between about 0.1-6mmol and oxo-proline between about 0.1-8 mmol, and/or L-lysine in anamount between about 0.1-12 mmol. n-acetyl L-cysteine and/or L-glutaminemay be comprised in some exemplary supplements at a level between about0.001-6 mmol. In particular examples, a functional dosage includesL-arginine at a level between about 2.5-4.5 mmol and oxo-proline betweenabout 4-6 mmol, and/or L-lysine in an amount between about 7-9 mmol.N-acetyl L-cysteine and/or L-glutamine may be comprised in someexemplary supplements at a level between about 0.001-0.5 mmol.

Certain embodiments herein include an oral nutritional supplement dosageform that consists of 0.86 mmol L-arginine; 1.32 mmol Oxo-proline; 2.05mmol L-lysine; 1.53 μmol N-acetyl L-cysteine; 1.71 μmol L-glutamine; and125 μg Schizonepta (aerial parts) powder (SeroVital™). Thus, an oralnutritional supplement dosage form may consist of 181.38 mg L-arginineHCl; 170.93 mg L-pyroglutamic acid; 374.83 mg L-lysine HCl; 0.25 mgN-acetyl L-cysteine USP; 0.25 mg L-glutamine; and 0.125 mg Schizonepta(aerial parts) powder, for example, in a capsule. In some examples, theoral nutritional supplement may be administered to a human being byorally administering 4 such dosage forms (i.e., 725.50 mg L-arginineHCl; 683.70 mg L-pyroglutamic acid; 1499.30 mg L-lysine HCl; 1.00 mgN-acetyl L-cysteine USP; 1.00 mg L-glutamine; and 0.50 mg Schizonepta(aerial parts) powder).

Some embodiments herein provide a method for increasing human growthhormone and/or testosterone in humans that comprises orallyadministering a nutritional supplement for jointly elevating growthhormone release and testosterone release to a healthy human being. Asused herein, a “healthy human being” refers to a human being having anyage-related decline in hGH and/or testosterone, excluding anyphysiological deficiency that is not age-related. Particular embodimentsinclude oral administration of a nutritional supplement for jointlyelevating growth hormone release and testosterone release to a humanthat is at least 30 years old.

A nutritional supplement for jointly elevating growth hormone releaseand testosterone release may be orally administered to a human being toimprove health, including by: increasing lean body mass; reducingobesity, adipose tissue mass, and anxiety; supporting weight loss;decreasing appetite and atrophic processes in skeletal muscle, liver,kidney, spleen, skin, and bone; and improving energy, endurance, sleep,metabolism, heart rate, blood pressure, cardiovascular health,sympathetic nervous activity, thyroid response, glucose utilization,mental/cognitive function, reaction time, bone density, hair health andappearance, nail health and appearance, skin health and appearance, andlibido.

In certain examples, oral administration of a nutritional supplement forjointly elevating growth hormone release and testosterone release mayimprove hair and/or nail thickness, length, and/or growth. Oraladministration of a nutritional supplement for jointly elevating growthhormone release and testosterone release may improve wound healing,provide anti-aging skin properties, reduce wrinkles, dark spots,discolorations, dullness, sagging, laxity, and thinning, and may furtherimprove texture, luminosity, “lift,” tone, radiance, smoothness,uniformity, and youthful look of skin.

A nutritional supplement for jointly elevating growth hormone releaseand testosterone release may be orally administered to a human being toimprove health by affecting one or more condition(s) and/or disease(s)that depend upon androgen activity. Such conditions and diseasesinclude, for example and without limitation: maintenance of musclestrength and function; reversal or prevention of frailty or age-relatedfunctional decline (“ARFD”) in the elderly (e.g., sarcopenia);remediation of age-related decreased testosterone levels in men;treatment of male menopause; treatment of hypogonadism; male hormonereplacement; treatment of male and female sexual dysfunction (e.g.,erectile dysfunction, decreased sex drive, sexual well-being, decreasedlibido); male and female contraception; hair loss; Reaven's Syndrome;enhancement of bone and muscle performance/strength; treatment ofcatabolic side effects of glucocorticoids; prevention and/or treatmentof reduced bone mass, density, and/or growth (e.g., osteoporosis andosteopenia); treatment of chronic fatigue syndrome (CFS); chronicmyalgia; treatment of acute fatigue syndrome and muscle loss followingelective surgery; acceleration of wound healing; acceleration of bonefracture repair; prevention of post-surgical adhesion formation;acceleration of tooth repair or growth; maintenance of sensory function(e.g., hearing, sight, olfaction and taste); treatment of periodontaldisease; treatment of wasting secondary to fractures and wasting inconnection with, for example, chronic obstructive pulmonary disease(COPD), chronic liver disease, AIDS, weightlessness, cancer cachexia,burn and trauma recovery, chronic catabolic state, eating disorders, andchemotherapy; treatment of cardiomyopathy; treatment ofthrombocytopenia; treatment of one or more symptoms of Crohn's disease;treatment of short bowel syndrome; treatment of irritable bowelsyndrome; treatment of inflammatory bowel disease; treatment ofulcerative colitis; treatment of complications associated withtransplantation; treatment of physiological short stature; treatment ofobesity and growth retardation associated with obesity; treatment ofanorexia (e.g., associated with cachexia or aging); treatment ofhypercortisolism and Cushing's syndrome; treatment of Paget's disease;treatment of osteoarthritis; induction of pulsatile growth hormonerelease; treatment of osteochondrodysplasias; treatment of depression,nervousness, irritability, and stress; treatment of reduced mentalenergy and low self-esteem (e.g., motivation/assertiveness); improvementof cognitive function (e.g., treatment of dementia, includingAlzheimer's disease and short term memory loss); treatment of catabolismin connection with pulmonary dysfunction and ventilator dependency;treatment of cardiac dysfunction (e.g., associated with valvulardisease, myocardial infarction, cardiac hypertrophy or congestive heartfailure); lowering of blood pressure; protection against ventriculardysfunction or prevention of reperfusion events; treatment of adults inchronic dialysis; reversal or slowing of the catabolic state of aging;attenuation or reversal of protein catabolic responses following trauma(e.g., reversal of the catabolic state associated with surgery,congestive heart failure, cardiac myopathy, burns, cancer, COPD etc.);reduction of cachexia and protein loss due to chronic illness such ascancer or AIDS; treatment of hyperinsulinemia, includingnesidioblastosis; treatment of immunosuppressed patients; treatment ofwasting in connection with multiple sclerosis or otherneuro-degenerative disorders; promotion of myelin repair; maintenance ofskin thickness; treatment of metabolic homeostasis and renal homeostasis(e.g., in the frail elderly); stimulation of osteoblasts, boneremodeling and cartilage growth; regulation of food intake; treatment ofinsulin resistance, including NIDDM; treatment of insulin resistance inthe heart; improvement of sleep quality and correction of the relativehyposomatotropism of senescence due to high increase in REM sleep and adecrease in REM latency; treatment of hypothermia; treatment ofcongestive heart failure; treatment of lipodystrophy; treatment ofmuscular atrophy; treatment of musculoskeletal impairment; improvementof the overall pulmonary function; treatment of sleep disorders; and thetreatment of the catabolic state of prolonged critical illness;treatment of hirsutism, acne, seborrhea, androgenic alopecia, anemia,hyperpilosity, benign prostate hypertrophy, adenomas and neoplasias ofthe prostate (e.g., advanced metastatic prostate cancer) and malignanttumor cells including the androgen receptor, such as is the case forbreast, brain, skin, ovarian, bladder, lymphatic, liver and kidneycancers; treatment of cancers of the skin, pancreas, endometrium, lungand colon; osteosarcoma; hypercalcemia of malignancy; metastatic bonedisease; treatment of spermatogenesis, endometriosis and polycysticovary syndrome; counteracting preeclampsia, eclampsia of pregnancy andpreterm labor; treatment of premenstrual syndrome; and treatment ofvaginal dryness.

A nutritional supplement for jointly elevating growth hormone releaseand testosterone release may be used in combination with other hormoneand/or steroid modulating supplements to enhance the effect of thenutritional supplements disclosed herein.

In accordance with the “consist essentially of” and “consistingessentially of” language herein, a nutritional supplement for jointlyelevating growth hormone release and testosterone release in someembodiments is essentially limited to the aforementioned ingredients,and does not include any additional active ingredients intended to addnutritional content (e.g., vitamins, minerals, etc.), but may includeadditional ingredients not intended to add nutritional content, forexample, ingredients intended to fulfill a non-nutritional purpose(e.g., coloring, fillers, flavoring, an ingredient for maintaining thestructural form, etc.).

Each ingredient of a nutritional supplement for jointly elevating growthhormone release and testosterone release may be prepared in accordancewith any method known to one of ordinary skill in the art.Alternatively, each ingredient may be obtained in a fully-prepared froma commercially available source.

A nutritional supplement for jointly elevating growth hormone releaseand testosterone release may be in any suitable oral administrationform, including but not limited to: a chewable form, a liquid form, aspray form, a capsule form, a suppository form, dissolvable wafer, and apowder form. In some embodiments, a dosage form of the nutritionalsupplement may be present in an amount of about 2.9 grams.

Irrespective of the structural form of the nutritional supplement forjointly elevating growth hormone release and testosterone release, theingredients of the nutritional supplement may be distributedhomogeneously or non-homogeneously within the nutritional supplement.

A nutritional supplement for jointly elevating growth hormone releaseand testosterone release may be ingested on a regular basis, such as adaily or weekly intake at a dosage tailored to an individual's needs;i.e., the nutritional supplement is to be taken regularly as multiples(1×, 2×, etc.) of the dosage form (e.g., pills, tablets, capsules, etc.)in accordance with the needs of the individual. For example, a seniorcitizen leading a sedentary life is likely to need higher daily dosesthan does a young person engaged in regular strenuous exercise (e.g., aweight lifter).

Alternatively, the nutritional supplement for jointly elevating growthhormone release and testosterone release may be ingested on an as-neededbasis at a dosage tailored to the individual's needs. Medical ornutritional counseling may be beneficial for arriving at a desirable oroptimal dosage tailored to the individual's needs. The nutritionalsupplement may be administered, for example, from one to three timesdaily, or, by way of further example, the supplement may be administeredevery other day, or once a week. In particular embodiments, thenutritional supplement may be administered on an empty stomach.

In certain embodiments, a nutritional supplement for jointly elevatinggrowth hormone release and testosterone release comprises a particularcombination of types of amino acids, mass ranges, and specificformulations that have been selected to be synergistically-balanced andof adequate quantity to achieve a desired physiological effect; e.g.,growth hormone release and testosterone release. Improper combinationsof the same amino acids may be ineffective. The component amino acidsmay be synergistic in the sense that several of them, when combinedtogether, synergistically stimulate the release of both human growthhormone and testosterone. The combination of amino acids in particularembodiments was also chosen to reduce or inhibit chemical combination orreaction between the component amino acids.

EXAMPLES Example 1 Effect of an Oral Nutritional Supplement Single Doseon hGH Levels

The short-term effects of a single oral nutritional supplement on hGHlevels 2 hours post ingestion was studied 16 healthy subjects [12 males,4 females; 9 Caucasian, 6 African American, 1 other; mean age=32±14years; body mass index=26.4±5.0 ranging from 19.1 to 36.8 kg/m²]. Eachsubject reported to the Inpatient Unit on two occasions one week apart.After an overnight fast, subjects had an IV line placed and baselinebloods samples were drawn at −30, −15, and 0 minutes.

Subjects were then asked to swallow the capsules of supplement(SeroVital™) or an identical looking placebo. SeroVital™ is a 2.9 g/doseblend of L-lysine HCl, L-arginine HCL, oxo-proline, N-acetyl-L-cysteine,L-glutamine, and schizonepeta (aerial parts) powder. Blood was drawn at15, 30, 60 and 90 and 120 minutes for assay. Human GH was measured ateach time point using the Siemens Immulite™ 2000 (intra-assay CV was3.72%, inter-assay CV was 5.70%, and the detection limit for GH was 0.05ng/mL. The −15 and 120 minute time points were additionally assayed fortriiodothyronine (T3) as informative for mechanistic investigations.

The mean growth hormone increased 682% after the supplement from 0.17 atbaseline to 1.33 ng/mL at 120 minutes, compared to a mean decrease of52% after the placebo from 0.93 to 0.45 ng/mL. FIG. 1.

The mean change in GH levels from baseline to 120 minutes (GH at 120minutes minus GH at 0 minutes), was 1.15 (95% CI: 0.17, 2.14) ng/mLafter the supplement, versus −0.48 (−1.47, 0.50) ng/mL after theplacebo, demonstrating a statistically-significant differential effect(P=0.01). After the supplement, the mean AUC for GH across 120 minuteswas 20.43 (95% CI: 19.90, 20.95) ng/mL/min which was significantlyhigher (P=0.04) than placebo at 19.67 (18.74, 20.59) ng/mL/min. Overall,120 minutes after taking the supplement, GH levels were significantlyhigher in absolute levels or by AUC.

As daily circadian levels of T3 naturally decrease during the morninghours, at which the current trial was scheduled, it was not surprisingthat placebo levels between the −15 and 120 minute time points decreasedby −6.10 ng/dL (106 to 100 ng/dL, P=0.01). In contrast, the SeroVital™group exhibited a deceased reduction in T3 by nearly one-half over thesame time course, −3.3 ng/dL (101-97.3 ng/dL, NS), which was not asignificant reduction compared to baseline, as was the reduction in theplacebo group. These results affirm that somatostatin inhibition plays amechanistic role in the ability of SeroVital™ to induce significantincreases in serum GH levels in human subjects.

At 120 minutes, GH concentrations were 2-fold higher in women (2.3±1.1ng/mL, n=4) than in men (1.0±0.4 ng/mL, n=12, although the study was notadequately powered for these comparisons. Nevertheless, these findingssupport an enhanced effect of the SeroVital™ supplement in women.

An eight-fold increase was observed, equivalent to 682%, in GH levels120 minutes after a single oral supplement of SeroVital™. The study hada broad range of ages and BMI's and included both genders. An additionaladvantage of the present study over previous GH evaluations is that itcontained a placebo control group and was randomized and double-blinded.

These findings demonstrate that a specialized low-dose amino acidsupplement can significantly increase short-term GH levels. Futurestudies will examine whether such increases in GH with oral amino acidsupplementation increase fat-free mass and strength. This indeed may bethe case, since elderly subjects administered oral GH secretagogues for6 and 12 months have sustained increases in lean body mass and improvedphysical function.

The absolute magnitudes of these results are somewhat difficult todirectly correlate among past studies, as commercial GH assays usedifferent antibodies to target specific GH epitopes. Therefore,different antibodies and assays are less likely to recognize somespecific isoforms and fragments of the GH molecule. This results invariability of the normal range of the GH measurements in differentassays. Indeed, the same GH sample measured using different assays canvary 2-3 fold, limiting the ability to compare actual GH levels acrossstudies. Nevertheless, the mean levels of GH reached after thesubcutaneous injection of 0.06 IU of HGH in the treatment of GHdeficient subjects was 0.4 ng/mL, a value that was clearly in the rangeof values seen in our study with oral amino acids.

Findings obtained from a randomized, blinded, placebo-controlled studystrengthen the evidence that oral administration of amino acids, whencompounded properly, can increase GH serum levels, wherein SeroVital™administration showed an 8-fold increase, equivalent to a 682% increasein GH levels, 120 minutes after a single oral dose. In addition, weelucidate some mechanistic details for these significant GH increases asthrough somatostatin inhibition, supported by our results on the 120minute results on T3 levels in the same subjects.

Example 2 Effect of an Oral Nutritional Supplement on Endurance and FatMetabolism

To test the effect of the nutritional supplement on endurance and fatmetabolism, a double-blind clinical study was conducted involving 12healthy subjects [7 males, 5 females; mean age=31±6 years; body massindex=25.7±3.8 ranging from 20.3 to 32.2 kg/m²]. Each subject reportedto the Fitness Testing Facility (PEAK, University of Utah College ofHealth) after an overnight fast. Upon arrival, each subjects underwentstandard measurements of weight, height, body fat percent (Bod Pod), andresting metabolic rate (RMR, indirect calorimerty). Daily calorieexpenditure was estimated based on the additive evaluations of measuredRMR, estimated Lifestyle and Activity (defined as the number of caloriesburned performing daily activities including working, playing eating,etc), and estimated Exercise (defined as an estimate of the number ofcalories burned during exercise based on daily activity level).Following the baseline measurements, subjects then consumed a standardbreakfast (Egg McMuffin, 300 Calories; 12 g fat; 29 g carbohydrates; 18g protein). Subjects rested for a further 45 minutes to reach apost-absorptive state, then underwent a Maximal Aerobic Fitness Test ofgraded exercise, completed on a treadmill. Subjects' oxygen uptake wasmeasured using a metabolic cart, and VO_(2max) was quantified.

Subjects were then provided a two week supply of a novel supplementSeroVital™ (2.9 g/dose blend of L-lysine HCl, L-arginine HCl,oxo-proline, N-acetyl-L-cysteine, L-glutamine, and schizonepeta (aerialparts) powder). The novel SeroVital™ increases serum human growthhormone hGH levels by 8 times (equivalent to 682%) 120 minutes after asingle dose in healthy male and female volunteers. In this study,subjects were instructed to consume one dose of the supplement on anempty stomach, two hours after dinner prior to bedtime, every night forthe two-week study duration.

Following the final dose, each subject returned to the PEAK FitnessTesting Facility, University of Utah College of Health after anovernight fast (without having consumed their last dose of thesupplement since the previous night). Upon arrival, each subjectsunderwent the identical test protocols as the baseline test day. Thedata from the two measurement days were then analyzed. Consistent withthe hypothesis that the supplement would increase improve enduranceparameters with its ability to increase in hGH levels, the decision wasmade to reject the null only if the data supported the one directionalalternative consistent with a favorable response to the supplement.Statistical significance was assumed for P≦0.05.

Mean VO_(2max) increased by 6% from 44.9±8.1 at baseline to 47.7±9.2mL/kg/min (3.69±0.96 to 3.91±1.02 L/min), demonstrating a statisticallysignificant differential effect compared to baseline (P=0.02). After theperiod of supplementation with SeroVital™, the mean RMR increased by2.7% from 1687±330 to 1733±288 kcal/day with a statistical trend towardssignificance compared to baseline (P=0.165). Estimated daily calorieexpenditure also increased by 2.7% from 1687±330 to 1733±288 kcal/daywith the trend towards significance (P=0.166).

After two weeks of supplementation with the supplement SeroVital™,(taken on an empty stomach, two hours after dinner prior to bedtime,every night), both RMR and estimated daily calorie expenditure tended toincrease, evidencing the potential of the supplement to impart long-termfat burning effects. Additionally, endurance as measured by VO_(2max) inthe post-absorptive state significantly improved with a measured 6%increase. Overall, the SeroVital™ supplement was shown to increaseparameters of endurance, energy, and vitality.

Example 3 Effect of an Oral Nutritional Supplement on Sleep Improvement

To test the ability of the nutritional supplement to improve sleep, adouble-blind clinical study was conducted involving 15 healthy subjects[10 males, 5 females; mean age=33±7 years]. Each subject completed abaseline Epworth Sleepiness Scale self-report questionnaire and astandardized assay of usual sleep habits. All subjects were deemed tohave average sleep parameters within a normal range.

The subjects were then provided a three week supply of a novelsupplement SeroVital™ (2.9 g/dose blend of L-lysine HCl, L-arginine HCl,oxo-proline, N-acetyl-L-cysteine, L-glutamine, and schizonepeta (aerialparts) powder). The novel SeroVital™ blend increases serum human growthhormone hGH levels by 8 times (equivalent to 682%) 120 minutes after asingle dose in healthy male and female volunteers.

We investigated sleep patterns with continued use of the supplement whentaken on an empty stomach, two hours after dinner prior to bedtime,every night for three weeks. On each trial day, subjects reported 1)time went to bed; 2) time of final wakening; 3) estimated time to fallasleep; 4) time of awakening during sleep/length of time awake. Data wascompiled by day for estimated time to fall asleep and length of timeawake during sleep in order to assess sleep efficiency. Daily values foreach measure were plotted as an average (±S.D.) among the subjects overthe time course of the study, and a linear regression was tabulated toassess overall trends over time. All available data was included in theanalysis.

Linear regression analysis showed that both estimated time to fallasleep (FIG. 2) and time awake during sleep (FIG. 3) tended to decreaseover time with continued use of the supplement over the time course ofthe study. The time to fall asleep decreased with an average slope of−0.24 min/day, and the time awake during sleep decreased by an averageslope of −0.26 min/day. Overall, these results so a trend towardsgreater sleep efficiency by measurements of both time to fall asleep andtime awake during sleep, both with a quantified average decrease ofabout 0.25 min/day over three weeks with regular nighttime use of thenovel SeroVital™ supplement (when taken as directed, on an emptystomach, two hours after dinner prior to bedtime).

Example 4 Effect of an Oral Nutritional Supplement on Lean Body Mass andWeight Change

To test the effect of the nutritional supplement on lean body mass andweight change of humans, healthy men and women between the ages of 30 to80 years are recruited through advertisements. Standardized assessmentcriteria are used to select subjects at risk for functional decline(e.g., hand grip strength, habitual gait speed, etc.). Additionally,subjects are excluded for diabetes mellitus, use of anticoagulants,seizure disorder, cancer treatment within five years, poorly controlledhypertension, unstable or recent onset angina, myocardial infarctionwithin 6 months, cognitive impairment, depression, significantlimitations of lower extremity function, bradycardia, systolic bloodpressure <100 or >170 mm Hg, or orthostatic hypotension. Subjectsparticipating in strength training programs were also excluded.

Subjects are given daily doses of the nutritional supplement for 6months at different dosing concentrations of active ingredients, and onegroup is treated as a placebo group. The study includes a pre-screeningassessment, screening and baseline visits, and weekly visits over theplanned 6 months, where weight, body composition, and physicalperformance (including endurance) are determined, along with hormonaldata analysis. Weight, percent lean body mass, and percent fat body massare chosen as primary measures of body composition at the 6 monthanalysis period.

Example 5 Effect of an Oral Nutritional Supplement on Energy, BoneDensity, Skin Thickness, and Mass of Adipose Tissue

To test the effect of the nutritional supplement on energy, bonedensity, skin thickness, and mass of adipose tissue of healthy men over60 years old are recruited through advertisements. Standardizedassessment criteria are used to select subjects at risk for functionaldecline (e.g., hand grip strength, habitual gait speed, etc.). Entrycriteria includes body weight of 90 to 120 percent of the standard forage. Additionally, subjects are excluded for diabetes mellitus, use ofanticoagulants, seizure disorder, cancer treatment within five years,poorly controlled hypertension, unstable or recent onset angina,myocardial infarction within 6 months, cognitive impairment, depression,significant limitations of lower extremity function, bradycardia,systolic blood pressure <100 or >170 mm Hg, or orthostatic hypotension.Subjects participating in strength training programs were also excluded.

Subjects are given daily doses of the nutritional supplement for 6months at different dosing concentrations of active ingredients, and onegroup is treated as a placebo group. The study includes a pre-screeningassessment, screening and baseline visits, and weekly visits over theplanned 6 months, where lean body mass, mass of adipose tissue, skinthickness, and bone density at various skeletal sites are measured.Subjects are also monitored for changes in energy levels throughout theexperiment.

Example 6 Effect of an Oral Nutritional Supplement on Anxiety and Sleep

To test the effect of the nutritional supplement on anxiety and sleep,healthy men and women between the ages of 30 to 80 years are recruitedthrough advertisements. Subjects are interviewed by a psychiatrist usingthe Structured Clinical Interview for DSM-III-R or analogous method.Patients are all drug free. The study focuses on the evaluation ofanxiety, sleep patterns, and measurement of basal morning stress hormonelevels at various points during the 6 month testing period.

Subjects are given daily doses of the nutritional supplement for 6months at different dosing concentrations of active ingredients, and onegroup are treated as a placebo group. The study includes a pre-screeningassessment, screening and baseline visits, and weekly visits over theplanned 6 months, where anxiety assessment (e.g., Hamilton AnxietyRating Scale), and anxiety and stress hormone levels (e.g., cortisol andGH) are determined, and sleep study analysis are conducted.

Example 7 Effect of an Oral Nutritional Supplement on TestosteroneLevels

The effects of an oral nutritional supplement on testosterone levels arestudied in a group of healthy human subjects. Subjects are asked toswallow the capsules of SeroVital™, or an identical looking placebo.

Blood samples (8-10 mL) are taken from each participant before use,after 6 days of use, after 12 days of use, and 3 days after suspendinguse. Each blood draw is taken in the morning between 9:30 and 10:30 am,at a time when serum oscillations of testosterone is at its minimumdaily value. The determination of testosterone in human blood is carriedout using commercially available immunochemoluminescence assays. (Topoet al., 2009)

Results on testosterone release show that after using the SeroVital™supplement, serum testosterone levels are increased in a significantnumber of the subjects taking the supplement. On average, the increasein serum testosterone is statistically significant compared to those ofthe placebo group over the time course of the study.

While embodiments of the present invention have been described hereinfor purposes of illustration, many modifications and changes will becomeapparent to those skilled in the art. Accordingly, the appended claimsare intended to encompass all such modifications and changes as fallwithin the true spirit and scope of this invention.

We claim:
 1. A method for increasing sex steroids and human growthhormone in a human being in need thereof, the method comprising:providing an oral nutritional supplement unit dosage form, wherein theoral nutritional supplement unit dosage form comprises about 1 mmolL-arginine, about 1 mmol Oxo-proline, about 2 mmol L-lysine, about 1.5μmol N-acetyl L-cysteine, about 2 μmol L-glutamine, and about 125 μgSchizonepta (aerial parts) powder; and orally administering an effectiveamount of the nutritional supplement to the human being.
 2. The methodaccording to claim 1, wherein the oral nutritional supplement unitdosage form comprises: about 0.9 mmol L-arginine, about 1.3 mmolOxo-proline, about 2.0 mmol L-lysine, about 1.5 μmol N-acetylL-cysteine, about 1.7 μmol L-glutamine, and about 1.25 μg Schizonepta(aerial parts) powder.
 3. The method according to claim 2, wherein theoral nutritional supplement unit dosage form comprises about 181 mgL-arginine HCl; about 170 mg L-pyroglutamic acid; about 374 mg L-lysineHCl; about 0.25 mg N-acetyl L-cysteine USP; about 0.25 mg L-glutamine,and about 0.125 mg Schizonepta (aerial parts) powder.
 4. The methodaccording to claim 1, wherein the oral nutritional supplement unitdosage form is administered to the human being, so as to increase leanbody mass or libido; reduce obesity, adipose tissue mass, or anxiety;support weight loss; decrease appetite and atrophic processes inskeletal muscle, liver, kidney, spleen, skin, or bone; improve energy,endurance, sleep, metabolism, heart rate, blood pressure, cardiovascularhealth, sympathetic nervous activity, thyroid response, fat metabolism,glucose utilization, mental/cognitive function, reaction time, bonedensity, hair health and appearance, nail health and appearance, skinhealth and appearance, or libido; and/or affect one or more condition(s)and/or disease(s) that depend upon androgen activity in the human being.5. The method according to claim 1, wherein the oral nutritionalsupplement unit dosage form is orally administered in a total amount ofabout 2.9 grams.
 6. The method according to claim 1, wherein the oralnutritional supplement is administered from one to three times daily. 7.The method according to claim 1, wherein the oral nutritional supplementis administered once a week.
 8. The method according to claim 1, whereinthe human being is at least 30 years old.
 9. The method according toclaim 1, wherein the human being exhibits one or more symptoms ofpost-traumatic stress disorder selected from the group consisting ofre-experiencing the original trauma(s) through flashbacks or nightmares,avoidance of stimuli associated with the trauma, increased arousal,difficulty falling or staying asleep, anger, and hypervigilance.